Exogenous dsRNA trigger RNAi in Venturia inaequalis resulting in down regulation of target genes and growth reduction.

BACKGROUND: Venturia inaequalis is an apple scab causing fungal pathogen. It is a highly contagious and destructive pathogen which rapidly spreads infection in the surrounding orchards if not managed. The management and control of disease require multiple fungicides to be sprayed at different development stages of the apple. Persistent applications of fungicides also raises environmental concerns. Here, we demonstrate the potential of using spray induced gene silencing (SIGS) by developing target specific gene constructs for the synthesis of corresponding double-stranded RNA (dsRNA). METHODS AND RESULTS: The exogenous application of dsRNAs was found to reduce mycelial growth and spore formation of V. inaequalis on culture plates. Four genes of V. inaequalis viz. CIN1, CE5, VICE12 and VICE16 which get upregulated during infection, were selected as targets for the development of gene construct expressing the corresponding dsRNA. The effect of exogenously supplied in vitro synthesized dsRNA on V. inaequalis was assessed in culture bioassay experiments with respect to growth, and spore formation. The expression level of the target genes in treated and control fungus was evaluated using quantitative PCR. Fungus treated with VICE12 targeted dsRNA showed maximum reduction in colony size (~ 55%), conidia formation (~ 93%) and expression level of the corresponding gene (2.2 fold), which was followed by CIN1-dsRNA. VICE16-dsRNA treatment was least effective with 32% reduction in growth, the non-significant effect of conidial spore formation and 1.13 fold down regulation of corresponding target gene expression level. CONCLUSION: The result of this investigation validates the hypothesis that RNAi is evoked in V. inaequalis by exogenously supplied dsRNA and spray induced gene silencing (SIGS) based solutions may reduce burden of fungicide usage on apple crop against apple scab disease in future.

A waste-based circular economy approach for phycoremediation of X-ray developer solution.

A waste-based circular economy approach is proposed for the phycoremediation of an X-ray developer (XD) solution. The present study emphasizes the utilization of food waste (FW) and agri-compost media (ACM) as growth media for D. armatus for the subsequent bioremediation potential of XD solution-coupled lipid production. A 3:1 dilution (FW/ACM: XD.) was found to be suitable for the phycoremediation study of XD solution towards the % removal of biological oxygen demand (BOD), chemical oxygen demand (COD) and silver. The phycoremediation studies of diluted XD solution in FW demonstrated a 74.50% BOD removal, 81.69% COD removal, and 54.70% removal of silver. The growth of D. armatus in diluted XD solution in food waste was 1.37% lipid content. The phycoremediation of diluted XD solution with ACM resulted in 83.05% BOD removal, 88.88% COD removal and 56.30% silver removal with the concomitant lipid production of 1.42%. The optimal bioremediation coupled lipid production of D. armatus was observed on the 19th day of D. armatus cultivation in the developer effluent, along with food waste and agri-compost media, for 31 days. The study suggests a sustainable utilization of waste (FW and ACM) as a nutritive medium to scrutinize the phycoremediation of XD solution with a concomitant lipid production that can open up new avenues in phycoremediation coupled energy commodities production.

Functional foods as a formulation ingredients in beverages: technological advancements and constraints.

As a consequence of expanded science and technical research, the market perception of consumers has shifted from standard traditional to valuable foods, which are furthermore nutritional as well as healthier in today's world. This food concept, precisely referred to as functional, focuses on including probiotics, which enhance immune system activity, cognitive response, and overall health. This review primarily focuses on functional foods as functional additives in beverages and other food items that can regulate the human immune system and avert any possibility of contracting the infection. Many safety concerns must be resolved during their administration. Functional foods must have an adequate amount of specific probiotic strain(s) during their use and storage, as good viability is needed for optimum functionality of the probiotic. Thus, when developing novel functional food-based formulations, choosing a strain with strong technological properties is crucial. The present review focused on probiotics as an active ingredient in different beverage formulations and the exerting mechanism of action and fate of probiotics in the human body. Moreover, a comprehensive overview of the regulative and safety issues of probiotics-based foods and beverages formulations.

Effect of Salinity Stress on Lipid Accumulation in Scenedesmus sp. and Chlorella sp.: Feasibility of Stepwise Culturing.

The enhanced lipid accumulation in microalgae is envisioned under special stress conditions with the cost of algal growth, which in turn affects the overall lipid productivity. The selection of suitable stress conditions facilitates better lipid productivity without any harmful effect on microalgae growth and algal biomass production. In the present study, we have attempted to select the best salinity conditions towards better growth, biomass accumulation, and lipid productivity of microalgae. The study also envisaged testing the feasibility of the stepwise salinity stress-induced cultivation approach to minimize the growth penalty effect of microalgae. The highest specific growth rate (0.129, 0.133, 0.113 µday-1) and doubling per day (0.185, 0.193, 0.163 per day) were obtained at salinity concentration of 40 mM NaCl in BG-11 medium for Scenedesmus quadricauda (Sq19), Scenedesmus dimorphus (Sd12), and Chlorella sp. (Chl16), respectively. Maximal lipid content of 18.28, 30.70, and 32.19%, and lipid productivity of 8.59, 13.81, and 10.27 mg l-1 day-1 were achieved at 160 mM of NaCl in BG-11 media with the Sq19, Sd12, and Chl16 algal isolates, respectively. The utilization of stepwise salinity stress (160 mM) induced cultivation of Sd12 algal isolate results in higher lipid content (39.42%) and slightly improved lipid productivity than the control (without any stress, 20.4% lipid content). The results indicate the feasibility of enhancing the lipid content and productivity through the salinity-induced stepwise cultivation strategy.

Comparative de novo transcriptome analysis of male and female Sea buckthorn.

Sea buckthorn is a dioecious medicinal plant found at high altitude. The plant has both male and female reproductive organs in separate individuals. In this article, whole transcriptome de novo assemblies of male and female flower bud samples were carried out using Illumina NextSeq 500 platform to determine the role of the genes involved in sex determination. Moreover, genes with differential expression in male and female transcriptomes were identified to understand the underlying sex determination mechanism. The current study showed 63,904 and 62,272 coding sequences (CDS) in female and male transcriptome data sets, respectively. 16,831 common CDS were screened out from both transcriptomes, out of which 625 were upregulated and 491 were found to be downregulated. To understand the potential regulatory roles of differentially expressed genes in metabolic networks and biosynthetic pathways: KEGG mapping, gene ontology, and co-expression network analysis were performed. Comparison with Flowering Interactive Database (FLOR-ID) resulted in eight differentially expressed genes viz. CHD3-type chromatin-remodeling factor PICKLE (PKL), phytochrome-associated serine/threonine-protein phosphatase (FYPP), protein TOPLESS (TPL), sensitive to freezing 6 (SFR6), lysine-specific histone demethylase 1 homolog 1 (LDL1), pre-mRNA-processing-splicing factor 8A (PRP8A), sucrose synthase 4 (SUS4), ubiquitin carboxyl-terminal hydrolase 12 (UBP12), known to be broadly involved in flowering, photoperiodism, embryo development, and cold response pathways. Male and female flower bud transcriptome data of Sea buckthorn may provide comprehensive information at genomic level for the identification of genetic regulation involved in sex determination.

Enhanced microalgal lipid production with media engineering of potassium nitrate as a nitrogen source.

Algal biofuels are far from a commercial reality due to the technical challenges associated with their growth and lipid extraction procedures. In this study, we investigated the effect of 4 different media and 5 different nitrogen sources at 5 levels on the growth, biomass and lipid productivity of Scenedesmus sp and Chlorella sp The hypothesis was that a nitrogen source can be identified that provides enough stress to accumulate lipids without compromising significantly on biomass and lipid productivity. A maximum specific growth rate and doubling per day have been observed with algal species using modified BG-11 medium. Among the tested nitrogen sources, 2.5 mM potassium nitrate as a nitrogen constituent of modified BG-11 medium resulted in higher lipid content and productivity in the case of S. dimorphus (29.15%, 15.449 mg L-1day-1). Another noteworthy outcome of the present study lies in the usage of a smaller amount of the nitrogen source, i.e., 2.5 mM, which is found to be 7 times less than the standard BG11 media (17.60 mM sodium nitrate).

Characterization and Screening of Native Scenedesmus sp. Isolates Suitable for Biofuel Feedstock.

In current study isolates of two native microalgae species were screened on the basis of growth kinetics and lipid accumulation potential. On the basis of data obtained on growth parameters and lipid accumulation, it is concluded that Scenedesmus dimorphus has better potential as biofuel feedstock. Two of the isolates of Scenedesmus dimorphus performed better than other isolates with respect to important growth parameters with lipid content of ~30% of dry biomass. Scenedesmus dimorphus was found to be more suitable as biodiesel feedstock candidate on the basis of cumulative occurrence of five important biodiesel fatty acids, relative occurrence of SFA (53.04%), MUFA (23.81%) and PUFA (19.69%), and more importantly that of oleic acid in its total lipids. The morphological observations using light and Scanning Electron Microscope and molecular characterization using amplified 18S rRNA gene sequences of microalgae species under study were also performed. Amplified 18S rRNA gene fragments of the microalgae species were sequenced, annotated at the NCBI website and phylogenetic analysis was done. We have published eight 18S rRNA gene sequences of microalgae species in NCBI GenBank.

Sex-Biased Temporal Gene Expression in Male and Female Floral Buds of Seabuckthorn (Hippophae rhamnoides).

Seabuckthorn is an economically important dioecious plant in which mechanism of sex determination is unknown. The study was conducted to identify seabuckthorn homologous genes involved in floral development which may have role in sex determination. Forty four putative Genes involved in sex determination (GISD) reported in model plants were shortlisted from literature survey, and twenty nine seabuckthorn homologous sequences were identified from available seabuckthorn genomic resources. Of these, 21 genes were found to differentially express in either male or female flower bud stages. HrCRY2 was significantly expressed in female flower buds only while HrCO had significant expression in male flowers only. Among the three male and female floral development stages (FDS), male stage II had significant expression of most of the GISD. Information on these sex-specific expressed genes will help in elucidating sex determination mechanism in seabuckthorn.

Association studies for agro-physiological and quality traits of triticale x bread wheat derivatives in relation to drought and cold stress.

Correlation coefficient analysis conducted on 22 triticale x bread wheat derivatives along with six checks to select true- breeding derivative(s) for future hybridization programme with tolerance to drought and cold stress conditions as well as better quality traits revealed significant correlation of grain yield with spikelets per spike, biological yield, harvest index, leaf area index. Interestingly, the grain yield and drought susceptibility index showed no association. However, with cold tolerance it showed significant positive correlation indicating the desirability of certain plant traits under cold stress. The grain yield exhibited no association with quality traits which might assist in the predictability of high yielding varieties with high protein, total sugars, reducing sugars and non-reducing sugars. Path coefficient analysis revealed that biological yield had the highest positive direct effect on grain yield followed by harvest index, specific leaf weight, stomatal number, 1000 grain weight, stomatal size, spikelets per spike and days to heading. Therefore, indirect selection for these plant traits in order should be exercised in selecting drought tolerant genotypes. Two genotypes (RL-124-2P2 and RL 111P2) were found to be drought and cold tolerant with high grain yield, spikes per plant, spikelets per spike and leaf area index.

The release dynamics of model drugs from the psyllium and N-hydroxymethylacrylamide based hydrogels.

In order to utilize the psyllium husk, a medicinally important natural polysaccharide, for developing the novel hydrogels for the controlled drug delivery device, we have prepared psyllium and N-hydroxymethylacrylamide based polymeric networks by using N,N'-methylenebisacrylamide (N,N'-MBAAm) as crosslinker. The polymeric networks thus formed were characterized with scanning electron micrography (SEM), FTIR and thermogravimetric analysis (TGA) techniques to study various structural aspects of the networks and also with the swelling response of the polymeric networks as a function of time, temperature, pH and [NaCl]. Equilibrium swelling has been observed to depend on both structural aspects of the polymers and environmental factors. Maximum P(s) 748.3 was observed at 13.0 x 10(-3)mol/L of [N,N'-MBAAm] in 0.5M NaOH solution. The release dynamics of model drugs (salicylic acid and tetracycline hydrochloride) from hydrogels has also been discussed, for the evaluation of the release mechanism and diffusion coefficients. The effect of pH on the release pattern of tetracycline has been studied by varying the pH of the release medium. In release medium of pH 7.4 buffer the release pattern of tetracycline drastically changes to the extent that mechanism of drug diffusion shifted from non-Fickian diffusion to Fickian diffusion. It has been observed that diffusion exponent "n" have 0.71, 0.67 and 0.52 values and gel characteristic constant 'k' have 1.552 x 10(-2), 2.291 x 10(-2) and 5.309 x 10(-2) values in distilled water, pH 2.2 buffer and pH 7.4 buffer, respectively, for tetracycline release. In solution of pH 7.4 buffer, the rate of polymer chain relaxation was more as compare to the rate of drug diffusion from these hydrogels and it follows Fick's law of diffusion. The value of the initial diffusion coefficient for the release of tetracycline hydrochloride was higher than the value of late time diffusion coefficient in each release medium indicating that in the start, the diffusion of drug from the polymeric matrix was fast as compare to the latter stages.

In vitro regeneration of Acacia catechu Willd. from callus and mature nodal explants--an improved method.

Callus was derived from cultured cotyledons on MS medium supplemented with 2,4-D (0.25 mg/l) and NAA (0.25 mg/l). Plantlets were regenerated from the callus and nodal explants on MS medium containing BAP (2.0 mg/l) and Kn (2.0 mg/l), and further multiplied on the same medium. Addition of adenine sulphate (25.0 mg/l), ascorbic acid (20.0 mg/l) and glutamine (150.0 mg/l) in the medium resulted in enhanced axillary branching. Multiple shoots formed after 6 weeks were separated and subcultured in the fresh medium of same composition. For rhizogenesis, microshoots of 2.0-2.5 cm length were dipped in sterilized IAA solution (10 mg/l) for 24 hr followed by transfer to half strength MS medium containing activated charcoal (0.02%) resulting in rooting (75%) within 8 weeks. The rooted plants were transferred to pots containing sterilized soil and sand mixture for hardening and 71% survival was recorded. Fifty true to type plantlets of A. catechu could be obtained within seven months of culture establishment.